The glycoprotein hormones (GpHs), luteinizing hormone (LH), follicle-stimulating hormone (FSH) and thyroid-stimulating hormone (TSH), regulate fertility and metabolism. Their receptors (GpHRs) are distinguished from most G protein-coupled receptors by the presence of large N-terminal extracellular hormone-binding domains (ECDs). Each ECD consists of nine leucine-rich repeats (LRRs), flanked by N-terminal and C-terminal cysteine-rich regions (NCR and CCR, respectively). The signal of specific hormone binding to the ECD is conveyed by its CCR to the heptahelical transmembrane region (TM). Mutations, activating or inactivating GpHR function, have been identified in hyperthyroidism, precocious puberty and pseudohermaphroditism; moreover, polymorphic variants of LH and FSH receptors have been shown to change the susceptibility to female infertility, breast cancer and risk of bone fractures.
AIMS : 1. Identify common hormone-contact sites and key hormone-selective determinants in LRRs of GpHRs, 2. Identify residues in the cysteine-rich regions of GpHRs important for GpH binding and for conveying the binding signal to the TM, and 3. Determine the three-dimensional structure of GpH-GpHR complexes. METHODS: Domain exchange/site-directed mutagenesis, expression, hormone binding and cAMP-dependent reporter activity as readouts, molecular modeling, ECD protein expression, protein purification, crystallization, crystal structure determination. OUTCOME: The results of the proposed studies will increase our knowledge of the molecular mechanisms by which GpHRs recognize the 'correct' GpH, as well as the step-wise propagation of the signal to the TM following ligand binding. These data, in combination with the mutant 'gain-of-function' GpHRs, will be molded into a general model of GpHR activation, essential for the development of better treatment protocols and the rational design of drugs that change receptor activity. Moreover, these data may yield a better understanding of the etiology of different pathological states known for some GpH-GpHR pairs.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
1R01DK069711-01
Application #
6855902
Study Section
Molecular and Cellular Endocrinology Study Section (MCE)
Program Officer
Malozowski, Saul N
Project Start
2005-01-01
Project End
2008-12-31
Budget Start
2005-01-01
Budget End
2005-12-31
Support Year
1
Fiscal Year
2005
Total Cost
$357,615
Indirect Cost
Name
University of Georgia
Department
Biochemistry
Type
Schools of Arts and Sciences
DUNS #
004315578
City
Athens
State
GA
Country
United States
Zip Code
30602
Chen, Tsuey-Ming; Czerwiec, Frank S; Puett, David (2016) Steroidogenesis and early response gene expression in MA-10 Leydig tumor cells following heterologous receptor down-regulation and cellular desensitization. Biochem Biophys Rep 5:305-312
Cui, Juan; Yin, Yanbin; Ma, Qin et al. (2015) Comprehensive characterization of the genomic alterations in human gastric cancer. Int J Cancer 137:86-95
Cui, Juan; Xu, Ying; Puett, David (2013) Microarray-based transcriptome profiling of ovarian cancer cells. Methods Mol Biol 1049:119-37
Cui, Juan; Miner, Brooke M; Eldredge, Joanna B et al. (2011) Regulation of gene expression in ovarian cancer cells by luteinizing hormone receptor expression and activation. BMC Cancer 11:280
Angelova, Krassimira; Felline, Angelo; Lee, Moon et al. (2011) Conserved amino acids participate in the structure networks deputed to intramolecular communication in the lutropin receptor. Cell Mol Life Sci 68:1227-39
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Hong, Celine S; Cui, Juan; Ni, Zhaohui et al. (2011) A computational method for prediction of excretory proteins and application to identification of gastric cancer markers in urine. PLoS One 6:e16875
Cui, Juan; Eldredge, Joanna B; Xu, Ying et al. (2011) MicroRNA expression and regulation in human ovarian carcinoma cells by luteinizing hormone. PLoS One 6:e21730
Puett, David; Angelova, Krassimira; da Costa, Marcelo Rocha et al. (2010) The luteinizing hormone receptor: insights into structure-function relationships and hormone-receptor-mediated changes in gene expression in ovarian cancer cells. Mol Cell Endocrinol 329:47-55
García-López, Angel; de Jonge, Hugo; Nóbrega, Rafael H et al. (2010) Studies in zebrafish reveal unusual cellular expression patterns of gonadotropin receptor messenger ribonucleic acids in the testis and unexpected functional differentiation of the gonadotropins. Endocrinology 151:2349-60

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